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Updated: Oct 10, 2026
A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model
Published on: February 6, 2018
Dendrobium huoshanense ameliorates spermatogenesis disorder in rats through modulating m6A methylation modification
Renjie Tao1, Xia Li1, Leyuan Ji1
1College of Life and Health, West Anhui University, Generic Technology Research Center for Anhui Traditional Chinese Medicine Industry, Lu'an, Anhui, 237012, PR China.
Ethnopharmacological Relevance:
Spermatogenesis disorder (SPD) has become a significant factor affecting male reproductive health. Dendrobium huoshanense (DH) is a rare and precious perennial orchid used in traditional Chinese medicine to nourish yin and tonify the kidney, but its mechanism in improving spermatogenic function remains unclear.
Aim Of The Study:
This study aims to elucidate the pharmacodynamic material basis and mechanism of action of DH in improving SPD.
Materials And Methods:
Male rats were divided into blank control, model, positive drug, and DH high, medium, and low dose groups. SPD model was induced by Tripterygium wilfordii glycosides (TWG). Testicular histopathology, fibrosis, apoptosis, and protein levels of METTL3, METTL14, METTL16, WTAP, ALKBH5, TLR4, NRF2, and GPX4 were assessed. m6A-seq, metabolomics, and transcriptomics were performed to analyze m6A modifications and methyl donor abundance. Active components were identified via high-resolution mass spectrometry (HRMS).
Results:
DH aqueous extract attenuated spermatogenic cell loss, seminiferous tubules basement membrane fibrosis and germ cell apoptosis in SPD model group, up-regulated the protein expression levels of METTL3, METTL14, METTL16, WTAP, ALKBH5, NRF2, and GPX4 in testicular tissues of the model group, promoted consumption of carnitine derivative methyl donors, and increased m6A modification of Sox family (Sox4, Sox9, Sox11, Sox13, etc.), Nrf2, and Gpx4 in germ cells. The active components identified in DH aqueous extract primarily included bibenzyl and flavonoid compounds.
Conclusion:
DH up-regulates the METTL3/METTL14 complex, promoting methyl donor consumption and enhancing m6A modification and protein expression of Sox9, Gpx4, and Nrf2, thereby alleviating oxidative stress and restoring impaired spermatogenesis.